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Biomedical subjects

J Kiefer

Publications and source records attributed to J Kiefer.

At least 19 recordsLinked to original sources
PubMed1997

Separation of PP2A core enzyme and holoenzyme with monoclonal antibodies against the regulatory A subunit: abundant expression of both forms in cells.

Protein phosphatase 2A (PP2A) holoenzyme is composed of a catalytic subunit, C, and two regulatory subunits, A and B. The A subunit is rod shaped and consists of 15 nonidentical repeats. According to our previous model, the B subunit binds to repeats 1 through 10 and the C subunit binds to repeats 11 through 15 of the A subunit. Another form of PP2A, core enzyme, is composed only of subunits A and C. It is generally believed that core enzyme does not exist in cells but is an artifact of enzyme purification. To study the structure and relative abundance of different forms of PP2A, we generated monoclonal antibodies against the native A subunit. Two antibodies, 5H4 and 1A12, recognized epitopes in repeat 1 near the N terminus and immunoprecipitated free A subunit and core enzyme but not holoenzyme. Another antibody, 6G3, recognized an epitope in repeat 15 at the C terminus and precipitated only the free A subunit. Monoclonal antibodies against a peptide corresponding to the N-terminal 11 amino acids of the A alpha subunit (designated 6F9) precipitated free A subunit, core enzyme, and holoenzyme. 6F9, but not 5H4, recognized holoenzymes containing either B, B', or B" subunits. These results demonstrate that B subunits from three unrelated gene families all bind to repeat 1 of the A subunit, and the results confirm and extend our model of the holoenzyme. By sequential immunoprecipitations with 5H4 or 1A12 followed by 6F9, core enzyme and holoenzyme in cytoplasmic extracts from 10T1/2 cells were completely separated and they exhibited the expected specificities towards phosphorylase a and retinoblastoma peptide as substrates. Quantitative analysis showed that under conditions which minimized proteolysis and dissociation of holoenzyme, core enzyme represented at least one-third of the total PP2A. We conclude that core enzyme is an abundant form in cells rather than an artifact of isolation. The biological implications of this finding are discussed.

Animals↗
PubMed1997

Optimization of channel number and stimulation rate for the fast continuous interleaved sampling strategy in the COMBI 40+.

OBJECTIVE: To investigate the interrelation between number of channels and stimulation rate in the continuous interleaved sampling strategy (CIS). SUBJECTS AND METHODS: Three of the first recipients of the new COMBI 40+ cochlear implant participated in consonant, vowel, number, and sentence tests. Speech understanding was evaluated for different combinations of number of active channels from two to twelve and stimulation rate per channel between 1,515 and 9,090 pulses per second. RESULTS: The results indicate that the optimum number of active channels is not necessarily the maximum number of usable channels.

Adult↗
PubMed1996

A follow-up study of long-term results after cochlear implantation in children and adolescents.

The time course of speech development in children after cochlear implantation may extend over many years, thus making long-term studies necessary to evaluate any outcome. We report our long-term results after cochlear implantation in children and adolescents. Mean follow-up was 28 months, ranging from 1 to 5 years. After at least 1 year of experience all children were found to benefit from their cochlear implants. The majority of children scored above chance in speech identification tasks requiring closed set word and sentence understanding). At the 4-year interval, all children tested including prelingually deaf children had developed open set sentence understanding. The most relevant factor accounting for differences in the results was the duration of implant use in all groups. Even beyond 3 years the results continued to improve. Peri- or postlingually deafened children tended to have favorable results. For prelingually deaf children, duration of deafness and age at implantation were correlated negatively with the results.

Adolescent↗
PubMed1996

Prescribing practice with cognition enhancers in outpatient care: are there differences regarding type of dementia?--Results of a representative survey in lower Saxony, Germany.

Previous studies of cognition enhancers have mainly focused on insufficiently defined groups of cognition disorders, e.g., "cerebral insufficiency". With regard to the various biological changes in senile dementia of Alzheimer's type (SDAT) and in vascular dementia (VD), which together make up the great majority of senile dementias, many authors have encouraged different studies of these types of dementias, especially since both can be diagnosed clinically with satisfying certainty. Since primary care physicians treat the majority of elderly and demented patients, they have their own experience with cognition enhancers. We were therefore interested to know, how far these physicians differ in their treatment of SDAT and VD. We performed a representative survey (response rate 83.2%; 145 family physicians and 14 neuropsychiatrists) in the Goettingen area. A written case vignette described a 70-year-old widow with moderate dementia and vascular risk factors which are easily treated with drugs. Two versions were randomly assigned, in which (version A) either a "typical" VD history or a typical SDAT history (version B) were described. After perusal, the physician was asked whether and which drugs he would choose to treat the cognitive disorders in this patient. Most frequently, piracetam (A/B: 25.6%/30.9%), ginkgo biloba (24.4%/28.4%), and nimodipine (14.1%/25.9%) were considered. Aspirin was cited by 29.5%(A) and 17.3%(B) of the physicians respectively. As far as the type of dementia was concerned, significant differences were found only for co-dergocrine, which was preferred in SDAT. The following inter-group trends were observed: family physicians considered ginkgo biloba more often than nimodipine or co-dergocrine. The results show the apparent importance of cost-and safety aspects, while the type of dementia has hardly any impact. The latter impression corresponds to the results of drug trials demonstrating no different efficacy. In our opinion, aspirin was not sufficiently taken into consideration.

Adult↗
PubMed1996

HPRT mutations in V79 Chinese hamster cells induced by accelerated Ni, Au and Pb ions.

Mutation induction by accelerated heavy ions to 6-TG resistance (HPRT system) in V79 Chinese hamster cells was investigated with Ni (6-630 Me V/u), Au (2.2, 8.7 Me V/u) and Pb ions (11.6-980 Me V/u) corresponding to a LET range between 180 and 12895 ke V/microns. Most experiments could only be performed once due to technical limitations using accelerator beam times. Survival curves were exponential, mutation induction curves linear with fluence. From their slopes inactivation- and mutation-induction cross-sections were derived. If they are plotted versus LET, single, ion-specific curves are obtained. It is shown that other parameters like ion energy and effective charge play an important role. In the case of Au and Pb ions the cross-sections follow a common line, since these ions have nearly the same atomic weight, so that they should have similar spatial ionization patterns in matter at the same energies. Calculated RBEs were higher for mutation induction than for killing for all LETs.

Animals↗
PubMed1995

Oxygen radical production and thiol depletion are required for Ca(2+)-mediated endogenous endonuclease activation in apoptotic thymocytes.

Glucocorticoid hormones stimulate apoptosis in thymocytes via a mechanism that involves changes in intracellular Ca2+, and exogenous Ca2+ can also directly promote the nuclear alterations of apoptosis (lamin degradation and chromatin cleavage) in isolated nuclei. Here we report that glucocorticoid treatment resulted in the production of reactive oxygen species and the depletion of reduced glutathione. Separation of apoptotic cells on Percoll gradients demonstrated that both effects selectively occurred in thymocytes undergoing apoptosis. Moreover, glucocorticoid-induced endonuclease activation was partially blocked by the antioxidant N-acetyl-L-cysteine. Although abrogation of methylprednisolone-induced Ca2+ increases using the intracellular Ca2+ buffer 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid resulted in inhibition of endonuclease activation, it failed to prevent GSH depletion. However, N-acetyl-L-cysteine almost completely blocked methylprednisolone-induced elevations in cytosolic calcium levels, indicating that oxidative stress was playing a role in the Ca2+ response. Our results support the idea that oxidative stress is a key component of the apoptotic effector pathway in thymocytes, and that it interacts, at least in part, with the Ca2+ response.

Acetylcysteine↗
PubMed1995

[Factors influencing the prescribing of nootropic drugs. Results of a representative inquiry in Lower Saxony].

AIM OF INVESTIGATION: To discover (1) to what extent patients' wishes and the extent of any abnormality of brain performance influence the frequency with which "nootropic" drugs (those thought to affect brain activity, e.g. piracetam, pyritinol, or improve cerebral circulation, e.g. xanthine derivatives, Ginkgo biloba, secale alkaloids, calcium antagonists) are prescribed; (2) the medical practitioner's expectations of the effectiveness of such medications. METHOD: In a personal interview, 145 family doctors and 14 neurologists in private practice in the Göttingen area of Germany (participation rate: 83.2% of those asked to participate) were questioned about fictitious cases (case 1: mild memory problem with or without expressed wish for medication; case 2: moderate dementia, of Alzheimer or multi-infarct type). The previously arranged interviews, which took place in the doctors' practice rooms, consisted of standardized open questions to the written case reports. RESULTS: Regardless of the wish of the patient and the extent and type of the abnormal brain function about 70% of all participating doctors would prescribe those drugs, even though about 56% had doubts about their effectiveness. About 28% expected a positive effect on brain performance. A nearly equal proportion of doctors would continue an existing drug regimen as would prescribe one. CONCLUSION: The prescription of the named group of drugs is influenced less by medical criteria than by factors which concern doctor-patient relationship.

Adult↗
PubMed1995

Functional glucocorticoid receptor expression is required for cAMP-mediated apoptosis in a human leukemic T cell line.

The involvement of the glucocorticoid receptor (GR) in cAMP-induced apoptosis in a GR-deficient derivative of the CEM.C7 human T-ALL line was investigated. Incubation of the parental CEM.C7 cells with agents that elevate cAMP levels (dibutyryl cAMP and forskolin) resulted in DNA fragmentation characteristic of apoptotic cell death, whereas the GR-deficient ICR.27 cells were insensitive to the cytolytic effects of cAMP. Reconstitution of GR expression by transfection not only restored glucocorticoid sensitivity to the ICR.27 cells, but also promoted sensitivity to induction of apoptosis by cAMP. Thus, cAMP-induced apoptosis in T cells appears to occur via ligand-independent stimulation of at least some aspects of glucocorticoid receptor function.

Apoptosis↗
PubMed1995

Degradation of lamin B1 precedes oligonucleosomal DNA fragmentation in apoptotic thymocytes and isolated thymocyte nuclei.

Chromatin condensation and nuclear envelope breakdown are characteristic features of apoptotic cell death, but the mechanisms underlying these phenomena have not been identified. Solubilization of nuclear lamin is responsible for both events in mitosis. In this work, we report that glucocorticoids stimulate rapid degradation of lamin B1 that occurs before oligonucleosomal DNA fragmentation in apoptotic thymocytes. Protease inhibitors and the Ca2+ buffering agent BAPTA-AM block lamin degradation and DNA fragmentation, indicating that the processes are regulated by similar or identical mechanisms. Incubation of isolated thymocyte nuclei with Ca2+ stimulates lamin degradation before the detection of oligonucleosomal DNA fragments. However, in contrast to lamin dissolution during mitosis and some other forms of apoptosis, glucocorticoid-induced degradation of lamin B1 in thymocytes is not accompanied by dephosphorylation-mediated activation of cdc2. Our results demonstrate that lamin degradation is an early feature of apoptosis in thymocytes and suggest that chromatin condensation and breakdown of the nuclear envelope may occur as a result of disruption of nuclear lamina architecture.

Animals↗
PubMed1995

Induction of HPRT- mutants in Chinese hamster V79 cells after heavy ion exposure.

The induction of resistance to 6-thioguanine by heavy ion exposure was investigated with various accelerated ions (oxygen-uranium) up to linear energy transfer (LET) values of about 15,000 keV/microns. Survival curves are exponential with fluence; mutation induction shows a linear dependence. Cross-sections (sigma i: inactivation, sigma m: mutation) were derived from the respective slopes. Generally, sigma i rises over the whole LET range, but separates into different declining curves for single ions with LET values above 200 keV/microns. Similar behaviour is seen for sigma m. The new SIS facility at GSI, Darmstadt, makes it possible to study the effects of ions with the same LET but very different energies and track structures. Experiments using nickel and oxygen ions (up to 400 MeV/u) showed that inactivation cross-sections do not depend very much on track structure, i.e. similar values are found with different ions at the same LET. This is not the case for mutation induction, where very energetic ions display considerably smaller induction cross-sections, compared with low-energy ions of identical LET. Preliminary analyses using the polymerase chain reaction (PCR) demonstrate that even heavy ions cause "small alterations" (small deletions or base changes). The proportion of the total deletions seems to increase with LET.

Animals↗
PubMed1995

Heavy ion-induced DNA double-strand breaks with yeast as a model system.

Cells of diploid yeast, Saccharomyces cerevisiae, were exposed to a variety of energetic heavy ions (provided by the UNILAC facility at the Gesellschaft für Schwerionenforschung, GSI), 241Am alpha-particles and 80-keV x-rays after which they were assessed for DNA double-strand breaks (DSB) using either the neutral sedimentation or the pulsed-field gel electrophoresis (PFGE) technique. Both yielded comparable results. The DSB production cross-sections are compared with inactivation studies performed for the same cells under identical conditions. The measurements show that with lighter ions DSB induction cross-sections increase with linear energy transfer (LET), but the situation is less clear with the heavier ions. A close parallelism was found between DSB induction and cell inactivation in these yeast cells.

Alpha Particles↗
PubMed1995

Ambulatory blood pressure monitoring in healthy schoolchildren.

Ambulatory blood pressure monitoring (ABPM) was performed in 564 healthy schoolchildren during normal circadian activities. The data of two cohorts (155 boys and 139 girls aged 9-13 years and 184 boys and 168 girls with a body height between 120 and 155 cm) are presented. From the age of 9 to 13 years the mean 24-h systolic/diastolic blood pressure (SBP/DBP) increases from 107 +/- 9/66 +/- 7 mmHg to 115 +/- 13/68 +/- 9 mmHg in boys and from 104 +/- 5/64 +/- 6 mmHg to 109 +/- 8/65 +/- 9 mmHg in girls. When related to body height the values rise from 105 +/- 6/64 +/- 6 mmHg at 120 cm to 113 +/- 8/67 +/- 7 mmHg at 155 cm in boys and from 100 +/- 7/65 +/- 7 mmHg to 112 +/- 9/66 +/- 9 mmHg in girls. In comparison with the causal blood pressure data obtained from European studies, the presented ABPM values (daytime BP) are higher throughout, which may be explained by the increased activity during daytime with ABPM. There is a mean difference of 4.4 mmHg in boys and of 3.0 mmHg in girls for SBP and of 10.8 mmHg in boys and of 9.0 mmHg in girls for DBP when related to age. In relation to body height, there is a mean difference of 4.4 mmHg in boys and of 3.5 mmHg in girls for SBP and of 10.9 mmHg in boys and of 10.5 mmHg in girls for DBP. We conclude that standards derived from causal blood pressure measurements should not be used for the evaluation of ABPM data.

Adolescent↗
PubMed1995

Sleep disturbances in the demented elderly: treatment in ambulatory care.

We report the results of a representative survey in Lower Saxony, Germany, that focused on the treatment of sleep disturbances in the moderately demented elderly. Two written sample case histories (vignettes) described either a vascular demented patient suffering from nocturnal wandering or an Alzheimer's-type demented patient without apparent psychotic or behavioral (sleep) disorder. These were randomly assigned and presented to 145 family physicians and 14 neuropsychiatrists working in private practice by a trained investigator, who then conducted a standardized interview with the physicians. The study was representative of physicians (response rate: 83.2%). In response to the question concerning how they would treat the patient's sleep disturbances, about 20% of the physicians (with respect to both versions) answered that they would not choose drugs. More than 40% considered neuroleptics to be the drugs of choice. Benzodiazepines, antidepressants and other substances were seldom considered. No significant difference was noted in the response to the two different case histories. The results allow for the conclusion that non-drug treatments, which (at least initially) should be the treatment of choice, are mainly disregarded by the majority of the ambulatory care physicians. The reason for this seems to be a lack of education in sleep medicine and also in geriatric medicine.

Aged↗
PubMed1995

Killing and mutation of Chinese hamster V79 cells exposed to accelerated oxygen and neon ions.

Mutation induction by accelerated heavy ions to 6-thioguanine resistance (HPRT system) in Chinese hamster V79 cells was investigated using oxygen and neon ions with energies between 1.9 and 400 MeV/mu, corresponding to LET values between 18 and 754 keV/microns, respectively. Because of technical limitations most experiments could be performed only once. Inactivation and mutation induction cross sections, sigma i and sigma m, were obtained from the slopes of the exponential survival and the linear mutation induction curves, respectively. Both parameters increased with LET up to about 200 keV/microns, where the curves separated for the two types of ions. Calculated RBEs were higher for mutation induction than for killing for all LET values.

Animals↗
PubMed1995

Reasons for prescribing cognition enhancers in primary care. Results of a representative survey in Lower Saxony, Germany.

With regard either to the controversial debate about the efficacy of cognition enhancers (CEs) or to the high costs which the frequent prescription of these drugs causes the German health system's economy, we wanted to know what physicians expect from a therapy with these drugs. We performed a representative survey (response rate 83.2%) in Lower Saxony, Germany from February to July 1993. We designed two written case vignettes which described either a patient with slight memory problems or a moderately demented patient who also suffers from common systemic disorders. In a face-to-face interview 145 general practitioners and primary care internists (family physicians) and 14 community neuropsychiatrists answered the question, whether they would prescribe CEs to each of the patients described and what they would expect from this therapy. 70.4% of all physicians would prescribe a cognition enhancer to the slightly impaired patient and 63.5% to the multimorbid moderately demented patient, respectively. More than 50% of the family physicians would not expect any positive therapeutic effect in both patients, while the neuropsychiatrists did so in 57.1% in the patient with slight memory disturbances and in 35.7% in the moderately demented patient. A positive effect on cognition was expected by 28.2% of all physicians in the slight and by 18.3% in the moderately impaired patient, respectively. Other reasons mentioned were amelioration of cerebral perfusion and drive, as well as effects on disease progression. In conclusion, the results of this study clearly demonstrate that cognition enhancers are prescribed in spite of major doubts in their efficacy.

Aged↗
PubMed1994

Isolation and chromosomal localization of a novel human G-protein-coupled receptor (GPR3) expressed predominantly in the central nervous system.

Degenerate oligonucleotide primers designed against known G-protein-coupled receptors were used in polymerase chain reaction amplification to isolate a novel receptor sequence (R4) from a rat insulinoma cell line and its human homolog (GPR3) from a human neuroblastoma cDNA library. The novel human receptor sequence is expressed in low abundance predominantly in the central nervous system and at low levels in the lung and kidney. The gene encoding GPR3 is intronless within the coding region, contains at least one intron in the 5'-untranslated region, and has been localized to chromosome 1p34.3. The activating ligand for the homologous receptors R4 and GPR3 is not known, but sequence similarity with the closely related orphan rat receptor R334 [FEBS Lett. 292:243 (1991)] suggests that R334 and the homologous receptors R4 and GPR3 probably represent two discrete molecular subtypes that interact with the same or closely related ligands.

Amino Acid Sequence↗